Request for a ThermoFisher Helios 5UC DualBeam

7.项目概要/摘要： 此申请是为购买ThermoFisher Helios 5 UC双光束聚焦离子束扫描提供资金 电子显微镜（FIB-SEM）系统。该工具是由一组NIMH资助的 研究者需要获取脑组织的三维（3D）体积， 纳米分辨率所要求的FIB-SEM系统将安装在 匹兹堡大学医学院（UPSOM）的转化神经科学项目（TNP）。 所要求的Helios 5 UC DualBeam FIB-SEM的安装代表了现有 共享专门用于脑组织成像的显微镜设施。 存在对允许在3D大体积电子显微镜（VEM）方法中进行成像的3D大体积电子显微镜（VEM）方法的迫切且普遍的需求。 研究询问大脑机制的基础上复杂的行为和他们的中断， 精神疾病3D体积电子显微镜（VEM）的出现解决了这一关键需求， 生成高度详细的单独分辨的突触的连接组图，以及突触下细胞器， 在大量密集的脑组织中。所要求的FIB-SEM系统允许自动化 和脑组织的定量VEM成像，避免了与其他VEM相关的许多限制 接近。尽管FIB-SEM在神经科学研究中的应用相对较新，但我们表明， 技术可以应用于死后人脑组织，以创建3D、信息丰富的体积数据集 可以进行定量分析因此，有了这项技术，我们可以询问罚款 神经系统的结构和精确的结构破坏导致大脑功能障碍。 NIMH资助的研究人员的压倒性反应也证明了对这种仪器的需求 在UPSOM。我们清楚地解释了所要求的FIB-SEM将如何推进当前的研究目标。 NIMH资助的项目以及这些资源如何对产生新的NIMH赠款申请非常宝贵。的 该提案中的研究人员在传统EM方法方面有数十年的经验，但这些方法 不能产生由FIB-SEM成像产生的数据的数量或质量。因此，有一个 此应用的主要原因：提供急需的，尖端的，高质量的自动化VEM成像 在UPSOM内NIMH支持的用户。

7. Project Summary/Abstract: This request is for funds to purchase a ThermoFisher Helios 5UC DualBeam Focused Ion Beam Scanning Electron Microscope (FIB-SEM) system. This instrument is being requested by a group of NIMH-funded investigators who have a significant need to acquire 3-dimensional (3D) volumes of brain tissue imaged at nanometer resolution. The requested FIB-SEM system will be housed in a dedicated facility within the Translational Neuroscience Program (TNP) at the University of Pittsburgh School of Medicine (UPSOM). Installation of the requested Helios 5 UC DualBeam FIB-SEM represents an important expansion of existing shared microscopy facilities that are specialized for, and dedicated to, brain tissue imaging. There is a pressing and prevalent need for 3D large volume electron microscopy (VEM) approaches that permit studies interrogating the basis of brain mechanisms underlying complex behaviors and their disruptions in psychiatric disorders. The advent of 3D volume electron microscopy (VEM) addresses this critical need by generating highly detailed connectomic maps of individually resolved synapses, and the sub-synaptic organelles within, across densely packed, large volumes of brain tissue. The requested FIB-SEM system permits automated and quantitative VEM imaging of brain tissue that avoids many of the limitations associated with other VEM approaches. Although FIB-SEM application to neuroscience research is relatively recent, we show that this technology can be applied to postmortem human brain tissue to create 3D, information-rich datasets of volumes of brain tissue neuropil ready for quantitative analysis. Thus, with this technology we can interrogate the fine structure of the nervous system and pinpoint structural disruptions contributing to brain dysfunction. The need for this instrument is also demonstrated by the overwhelming response from NIMH-funded researchers in the UPSOM. We clearly explain how the requested FIB-SEM will advance the research objectives of current NIMH-funded projects and how this resource is invaluable for generating new NIMH grant applications. The investigators in this proposal have decades of experience in traditional EM approaches, but these methods cannot generate the quantity, or the quality, of data generated by FIB-SEM imaging. Therefore, there is a single prime reason for this application: to provide critically needed, cutting-edge, high quality automated VEM imaging to NIMH supported users within the UPSOM.